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Papers In Press, published online ahead of print July 11, 2002
J. Biol. Chem, 10.1074/jbc.C200398200
Submitted on July 8, 2002
Revised on July 11, 2002
Accepted on July 10, 2002

INSL3/Leydig insulin-like peptide activates the LGR8 receptor important in testis descent

Jin Kumagai, Sheau Yu Hsu, Hirotaka Matsumi, Jae-Sook Roh, Ping Fu, John Wade, Ross A. Bathgate, and Aaron J. Hsueh

Gyn/Ob, Stanford University, Stanford, CA 94305-5317

Corresponding Author: Aaron.Hsueh{at}stanford.edu

Several orphan G protein-coupled receptors homologous to gonadotropin and thyrotropin receptors have recently been identified and named as LGR4-8. INSL3, also known as Leydig insulin-like peptide or relaxin-like factor, is a relaxin family member expressed in testis Leydig cells and ovarian theca and luteal cells. Male mice mutant for INSL3 exhibit cryptorchidism or defects in testis descent due to abnormal gubernaculum development whereas overexpression of INSL3 induces ovary descent in transgenic females. Because transgenic mice missing the LGR8 gene are also cryptorchid, INSL3 was tested as the ligand for LGR8. Here, we show that treatment with INSL3 stimulated cAMP production in cells expressing recombinant LGR8, but not LGR7. In addition, interactions between INSL3 and LGR8 were demonstrated following ligand receptor cross-linking. Northern blot analysis indicated that the LGR8 transcripts are expressed in gubernaculum whereas treatment of cultured gubernacular cells with INSL3 stimulated cAMP production and thymidine incorporation. The present study identified the ligand for an orphan GPCR based on common phenotypes of ligand and receptor null mice. Demonstration of INSL3 as the ligand for LGR8 facilitates understanding of the mechanism of testis descent and allows studies on the role of INSL3 in gonadal and other physiological processes.


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